text: FSL constructs, when in solution (saline) and in contact, will spontaneously incorporate into cell membranes. The methodology involves simply preparing a solution of FSL constructs in the range of 1–1000 μg/mL, with the concentration used determining the amount of antigen present on the kodecyte. The ability to control antigen levels on the outside of a kodecyte has allowed for manufacture of quality control sensitivity systems and serologic teaching kits incorporating the entire range of serologic agglutination reactions. The actual concentration will depend on the construct and the quantity of construct required in the membrane. One part of FSL solution is added to one part of cells (up to 100% suspension) and they are incubated at a set temperature within the range of 4–37 °C (39–99 °F) depending on temperature compatibility of the cells being modified. The higher the temperature, the faster the rate of FSL insertion into the membrane. For red blood cells incubation for 2 hours at 37 °C achieves >95% FSL insertion with at least 50% insertion being achieved within 20 minutes. In general, for carbohydrate based FSLs insertion into red blood cells, incubation for 4 hours at room temperature or 20 hours at 4 °C are similar to one hour at 37 °C . The resultant kodecytes do not required to be washed, however this option should be considered if an excess of FSL construct is used in the "koding process".
  Kodecytes can also be created in vivo by injection of constructs directly into the circulation. However this process will modify all cells in contact with the constructs and usually require significantly more construct than in vitro preparation, as FSL constructs will preferentially associate with free lipids.
  The in vivo creation of kodecytes is untargeted and FSL constructs will insert into all cells non-specifically, but may show a preference for some cell types.
  Diagnostic serological analyses including flow cytometry and scanning electron microscopy usually can't see a difference between "kodecytes" and unmodified cells. However, when compared with natural cells there does appear to be a difference between IgM and IgG antibody reactivities when the functional group (F) is a monomeric peptide antigen. IgM antibodies appear to react poorly with kodecytes made with FSL peptides. Furthermore, FSL constructs may have a restricted antigen/epitope and may not react with a monoclonal antibody unless the FSL construct and monoclonal antibody are complementary.
  Kodecytes can be studied using standard histological techniques. Kodecytes can be fixed after "koding" subject to the functional moiety (F) of the FSL construct being compatible with the fixative. However, freeze cut or formalin-fixed freeze cut tissues are required because the lipid based FSL constructs (and other glycolipids) will be leached from the "kodecytes" in paraffin imbedded samples during the deparaffination steps.